Action Potentials ; Animals ; Bufo bufo ; Caffeine ; pharmacology ; Female ; Heart ; drug effects ; physiology ; In Vitro Techniques ; Male ; Sciatic Nerve ; physiology

Objectives To compare the characteristics of myasthenia gravis (MG) with different antibodies.Methods The muscle specific receptor tyrosine kinase (MuSk) and acetylcholine receptor (AChR) antibodies were detected in the sera of 119 MG patients,and fluoroimmunoprecipitation assay and cell based assay were applied. The clinical features of AChR-Ab positive,MuSK-Ab positive and serum negative MG patients were compared.Results There were 90 AChR-Ab positive sera tested out from the 119 MG sera,and 5 sera found with MuSK-Ab in the 29 AChR-Ab negative sera.There was no significant difference among the three groups regarding sex and age at onset.There were 3/5 of MuSK-Ab positive patients with predominantly bulbar paralysis,2/5 of MuSK-Ab positive patients were classified as MGFA Ⅲ to Ⅴ,and 79.2% (19/24) of serum negative patients were classified as MGFA Ⅰ.There was significantly positive relation between the levels of MuSK antibodies and disease severity (r=0.941,P=0.014).Neither thymic hyperplasia nor hymoma were found in MuSK-Ab positive patients.Conclusions MuSK antibodies are only detected out in the sera without AChR-Ab.The MuSK-Ab positive patients are mainly involved bulbar muscles,and without thymus abnormality.MuSK-MG is different with the AChR MG.

OBJECTIVE:To establish the method for simultaneous determination of gallic acid,rosmarinic acid,liquiritin and ammonium glycyrrhetate in Regan saibisitan granules. METHODS:RP-HPLC method was adopted. The determination was per-formed on Waters RP-C18 column with mobile phase consisted of acetonitrile-0.2% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelengths were 210 nm(gallic acid,rosmarinic acid and liquiritin),230 nm(ammonium glyc-yrrhetate). The column temperature was 28 ℃,and sample size was 20 μL. RESULTS:The linear ranges were 0.2744-7.546 μg for gallic acid(r=0.9998),0.1870-5.143 μg for rosmarinic acid(r=0.9996),0.1300-3.575 μg for liquiritin(r=0.9999)and 0.2540-6.985μg for ammonium glycyrrhetate(r=0.9998),respectively. The LOQ were 2.67,1.36,1.09 and 2.11 ng,respective-ly. The LOD were 1.03,0.62,0.87 and 0.91 ng,respectively. RSDs of precision,stability and repeatability tests were all less than 2.0%. The average recoveries were 97.26%-101.00%(RSD=1.1%,n=9),97.66%-101.80%(RSD=1.3%,n=9),97.45%-101.70%(RSD=1.4%,n=9),97.74%-101.70%(RSD=1.4%,n=9). CONCLUSIONS:The method is simple,accurate and reproducible, and can be applied for simultaneous determination of gallic acid,rosmarinic acid,liquiritin and ammonium glycyrrhetate in Regan saibisitan granules.

Dendritic cells(DC) are specialized antigen-presenting cells that prime naive T cells to induce initial immune responses. The immature DC capture and process antigens in the periphery, then emigrate to lymphoid organs. There they complete their maturation by upregulating HLA-I, II molecules, costimulatory molecules (eg. CD80, CD86) and adhesive molecules (eg. CD50, CD54, CD58). More studies showed that in vitro only interferons type I (IFN-alpha, beta) accelerate DC maturation in a dose-dependent manner. The DC induced by IFN type I highly express HLA-A, B, C, HLA-DR, costimulatory molecules and adhesive molecules, and they express enhancing effect of T-cells stimulatory activity in vitro. Progress of research in this field was summarized in this paper.
Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; physiology ; Humans ; Immunophenotyping ; Interferon Type I ; pharmacology

This study was aimed to investigate the effect of dendritic cells (DC) on the proliferation capability, immunophenotype changes, level of secreted cytokines and activity against leukemia of cytokine-induced killer (CIK) cells in vitro. DCs and CIK cells were induced from peripheral blood mononuclear cells of healthy volunteers. They were co-cultured meanwhile CIK cells were cultured alone as controls. Increased number of cells were counted by trypan-blue staining; the killing activity was detected by MTT assay; immunophenotype changes were analyzed by flow cytometry; the IL-12 and INF-gamma levels of the cultured supernatants were detected by ELISA kits. The results showed that the proliferation capability of DC-CIK cells was significantly higher than that of CIK cells (p < 0.05). Under the same condition, the ratio of double positive cells such as CD3(+) CD8(+), CD3(+) CD56(+) in CIK cells was significantly enhanced by co-cultured with DC cells (p < 0.05). The levels of IL-12 and INF-gamma in cultured supernatants of DC-CIK cells increased noticeably on day 3 as compared with CIK cells cultured alone (p < 0.01, p < 0.05). Within the effector-target ratio range between 5:1 to 40:1, the activity of DC-CIK cells against leukemia cells were much higher than that of CIK cells (p < 0.05), and this effect showed a positive correlation with the effector-target ratio. It is concluded that the proliferation capability of DC-CIK cells, the level of their secreted cytokines and their activity against leukemia cells are significantly higher than those of CIK cells. This research may suggest an approach for clinical immunotherapy against leukemia with DC-CIK cells.
Cell Line, Tumor ; Cell Proliferation ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; immunology ; metabolism ; Dendritic Cells ; cytology ; immunology ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism

This study was aimed to investigate the effect of cord blood dendritic cells (DCs) on the in vitro proliferation capability, immunophenotype changes, level of secreted cytokines and activity against leukemia cells of the homologous cytokine-induced killer (CIK) cells. DCs and CIK cells were induced from cord blood mononuclear cells. They were co-cultured at the ratio of 1:5, and CIK cells from cord blood or DC-CIK cells from peripheral blood were cultured as controls. Immunophenotypic changes were analyzed by flow cytometry, increased number of cells were counted by trypan-blue staining, the killing activity to leukemia cells was assayed by MTT, the levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-12 (IL-12) in the cultured supernatant were detected by ELISA. The results showed that the proliferation capability of cord blood DC-CIK cells was significantly higher than that of cord blood CIK cells and peripheral blood DC-CIK cells (p < 0.05 and p < 0.05). Under the same condition, the rate of double positive cells with CD3(+)CD8(+) and CD3(+)CD56(+) in CIK cells was significantly enhanced by co-culture with cord blood DCs (p < 0.05). The level of IL-12, IFN-γ, and TNF-α in cultured supernatants of cord blood DC-CIK cells increased noticeably on day 3 as compared with CIK cells cultured alone (p < 0.01, p < 0.05, p < 0.05). Within the effector-target ratio range between 2.5:1 to 20:1, the activity of cord blood DC-CIK cells against all subtypes of acute leukemia cells was much higher than that of CIK cells (p < 0.05), and there was no significant difference among all subtypes of acute leukemia cells, which was the same with the killing effect of peripheral blood DC-CIK cells against leukemia cells. It is concluded that the proliferation capability and anti-leukemia effect of the homologous CIK cells can be enhanced by cord blood DCs. The proliferation capability of cord blood DC-CIK cells is stronger than that of peripheral blood DC-CIK cells, but there is no significant differences of cytotoxicity between DCs and CIK cells. As the cord blood is easily gained and does not easily cause a serious graft rejection, the DC-CIK cells should be clinically applied more extensively as novel immune therapy.
Cell Proliferation ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Leukemia ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

To study the influence of IFN-alpha on function of CML-DC cultured in vitro and expression of chemokine and its chemokine receptor, bone marrow mononuclear cells from 13 CML patients were cultured in the fetal calf serum culture system supplemented with rhSCF, rhFlt-3L for expansion system, and adding rhGM-CSF, rhTNF-alpha, rhIL-4, with or without rhIFN-alpha to induce DCs. After incubation for two weeks, the phenotypes of CML-DC were analyzed by direct immunofluorescence and flow cytometry. The concentration of MIP-3beta expressed by CML-DC in the supernatant were analyzed by ELISA. The proliferative ability of T cells from healthy volunteers stimulated by CML-DCs were measured by MTT assay. The results showed that expression of CD86, CD83, CD40, MHC-I class molecules, CCR7, the concentration of MIP-3beta expressed by CML-DC, and the proliferative ability of T cells stimulated by CML-DCs in IFN-alpha group were all significantly higher than that in control group (P < 0.01). It is concluded that the immunophenotype of CML-DCs can be partially changed by IFN-alpha to accelerate the maturation of CML-DCs, enhance the capacity of CML-DCs, and stimulate allogeneic T lymphocyte proliferation.
Adult ; Aged ; Antigens, CD ; analysis ; B7-2 Antigen ; analysis ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; CD40 Antigens ; analysis ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chemokines ; biosynthesis ; Dendritic Cells ; drug effects ; metabolism ; pathology ; Female ; Flow Cytometry ; Fluorescent Antibody Technique, Direct ; Humans ; Immunoglobulins ; analysis ; Interferon-alpha ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; Leukocytes, Mononuclear ; drug effects ; metabolism ; pathology ; Male ; Membrane Glycoproteins ; analysis ; Middle Aged ; Receptors, Chemokine ; biosynthesis

AIMTo modify the C-5 side chains of the oxazolidinone derivatives and evaluate their in vitro antibacterial activities preliminarily.

METHODSThe title compounds were synthesized in 9 - 12 steps with the starting material 3-fluoroaniline and their in vitro antibacterial activities were examined by using Mueller-Hinton broth dilution method.

RESULTSThirty new compounds were designed and synthesized, in which eighteen novel title compounds were prepared and their structures were confirmed by 1H NMR and ESI-MS. Eleven compounds showed antibacterial activities to a certain extent, among them compounds 7a, 9a and 11a displayed promising activity.

CONCLUSIONCompounds 7a, 9a and 11a were worth further studying.

Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Microbial Sensitivity Tests ; Molecular Structure ; Oxazolidinones ; chemical synthesis ; chemistry ; pharmacology ; Staphylococcus aureus ; drug effects ; Structure-Activity Relationship

OBJECTIVE: To establish a simple and effective technique for detecting haplotype and heteroplasmy of mtDNA, and investigate their frequencies in Chinese Han population. METHODS: The fragments from 29-290 nt of mtDNA HV II from peripheral leukocytes of 200 unrelated Wuhan Han individuals were analyzed by using PCR-DGGE technique. RESULTS: Seventeen haplotypes were found in the range of 29-290 nt, and the haplotype diversity (HD) was 0.8844. The heterogeneity was observed from 4 individuals, and its frequency was 2%. CONCLUSION PCR-DGGE is a simple, sensitive and effective technique in analyzing polymorphism and heteroplasmy of mtDNA, and can be used in forensic practice.
Asian People/genetics* ; China/ethnology* ; DNA, Mitochondrial/genetics* ; Electrophoresis, Polyacrylamide Gel/methods* ; Genetic Heterogeneity ; Genetics, Population ; Haplotypes ; Humans ; Mutation ; Polymerase Chain Reaction/methods* ; Sequence Analysis, DNA

Objective: To evaluate the efficiency and safety of low intensity conditional regimen for children with Fanconi anemia (FA) receiving allogenic hematopoietic stem cells transplantation (allo-HSCT). Methods: Four patients diagnosed as Fanconi anemia were enrolled in this study. One patient received HLA-identical sibling donor hematopoietic stem cell transplantation, two patients underwent unrelated donor matched (UD) HSCT, and one patient received unrelated cord blood transplantation. The conditional regimen consisted of Busulfan with low dose of cyclophosphamide. Results: All 4 cases succeeded in allo-HSCT. The median time for neutrophils engraftment was 11(9-15) day, median time to platelets (PLT) engraftment was 12 (8-28) day. One case occurred with grade I of aGVHD, 1 case with hemorrhagic cystitis. No patient happened with hepatic veno-occlusive disease (VOD). Conclusion: Low intensity of conditional regimen is efficient and safe which should be recommended for FA patients with HSCT.
Busulfan ; Fanconi Anemia ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation Conditioning